LOCUS       pET-29c(+).        5372 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Bacterial vector for expression of N-terminally S-tagged proteins 
            with a thrombin site. For other reading frames, use pET-29a(+) or 
            pET-29b(+).
ACCESSION   .
VERSION     .
KEYWORDS    pET-29c(+).
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5372)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5372)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5372
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(140..157)
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    158..203
                     /label=MCS
                     /note="MCS"
                     /note="multiple cloning site"
     CDS             complement(217..234)
                     /label=thrombin site
                     /note="thrombin recognition and cleavage site"
     CDS             complement(250..294)
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     CDS             complement(295..297)
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     RBS             complement(305..327)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(342..366)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(367..385)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        698..775
                     /label=lacI promoter
     CDS             776..1855
                     /label=lacI
                     /note="lac repressor"
     protein_bind    1871..1892
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             2667..2855
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     misc_feature    2960..3102
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3288..3876)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             3998..4810
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(4906..5361)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
      181 cgacggagct cgaattcgga tccacagata tcccatggaa ccgcgtggca ccagggtacc
      241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatatg
      301 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
      361 aattccccta tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac
      421 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
      481 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
      541 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
      601 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
      661 atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac
      721 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
      781 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
      841 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
      901 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
      961 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     1021 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     1081 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     1141 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     1201 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     1261 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     1321 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     1381 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     1441 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     1501 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     1561 gcgcgttggt gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     1621 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     1681 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     1741 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     1801 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     1861 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
     1921 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     1981 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     2041 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     2101 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     2161 tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc
     2221 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
     2281 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
     2341 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
     2401 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
     2461 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
     2521 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
     2581 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
     2641 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
     2701 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
     2761 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
     2821 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     2881 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2941 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     3001 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tatgcggtgt
     3061 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg
     3121 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3181 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3241 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3301 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3361 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3421 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3481 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3541 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3601 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3661 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3721 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3781 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3841 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3901 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gaacaataaa
     3961 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
     4021 gtcttgctct aggccgcgat taaattccaa catggatgct gatttatatg ggtataaatg
     4081 ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg ggaagcccga
     4141 tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg ttacagatga
     4201 gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca agcattttat
     4261 ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa cagcattcca
     4321 ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg cagtgttcct
     4381 gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg
     4441 tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg attttgatga
     4501 cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac ttttgccatt
     4561 ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta tttttgacga
     4621 ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc gataccagga
     4681 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
     4741 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
     4801 tgagtttttc taagaattaa ttcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4861 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgaaatt gtaaacgtta
     4921 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     4981 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     5041 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     5101 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     5161 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     5221 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     5281 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     5341 atgcgccgct acagggcgcg tcccattcgc ca
//
